l-glutaminases is an important industrial enzyme which finds potential applications in different sectors ranging from therapeutic to food industry. It is widely distributed in bacteria, actinomycetes, yeast and fungi. l-Glutaminases are mostly produced by Bacillus and Pseudomonas sp. and few reports were available with fungal, actinomycete and yeast system. Modern biotechnological tools help in the improved production as well as with tailor made properties for specific applications. Most of the genetic engineering studies were carried out for the production of l-glutaminase with improved thermo-tolerance and salt tolerance. Considering the potential of in vitro applications of l-glutaminase, extracellular enzymes are important and most microbes produce this enzyme intracellularly. Several research and developmental activities are going on for the extracellular production of l-glutaminase. This review discusses recent trends and developments and applications of l-glutaminases.